Method for the amplification of nucleic acids using heat transfer for nanoparticles

Abstract

A method for the amplification of nucleic acids, in which nanoparticles in a reaction volume transfer heat to their environment through excitation. The method comprises a step of providing nanoparticles with the nucleic acids in a reaction volume and one or more heating steps. In at least one of the heating steps, the heating is achieved at least partially through the excitation of the nanoparticles. The interval of the excitation is chosen to be shorter or equal to a critical excitation time.

The invention claimed is: 
     
       1. A method for the amplification of nucleic acids comprising amplifying the nucleic acids, wherein the amplification comprises one or more heating steps, and a step of providing nanoparticles with the nucleic acids in a reaction volume, wherein the heating in at least one heating step is achieved at least partially through excitation of the nanoparticles so that the nanoparticles transfer heat to their environment through the excitation, wherein through the excitation of the nanoparticles, the environment of the nanoparticles is heated locally, wherein the interval of the excitation is chosen to be shorter or equal to a critical excitation time t1=(s1*|x|) 2 /D, and wherein s1=100, |x| is the mean nanoparticle distance, and D is the thermal diffusivity of the medium between the nanoparticles. 
     
     
       2. The method according to  claim 1 , wherein the nucleic acids are amplified by a polymerase chain reaction. 
     
     
       3. The method according to  claim 1 , wherein the nanoparticles are excited by a laser. 
     
     
       4. The method according  claim 1 , wherein the nanoparticles are conjugated with oligonucleotides. 
     
     
       5. The method according  claim 4 , wherein one kind of conjugates of the nanoparticles and the oligonucleotides is conjugated with forward primers as well as reverse primers. 
     
     
       6. The method according to  claim 4 , wherein in the method, counter sequences are used, which can bind to the oligonucleotides, which have detached from the nanoparticles, to which the oligonucleotides had been attached previously. 
     
     
       7. The method according to  claim 4 , wherein filling molecules are attached to the nanoparticles. 
     
     
       8. The method according to  claim 4 , wherein the oligonucleotides on the nanoparticles contain a spacer sequence as a partial sequence. 
     
     
       9. The method according to  claim 4 , wherein the heat, which is transferred to the environment of the nanoparticles by the excitation of the nanoparticles, is sufficient to dehybridise the oligonucleotides on the surface of the nanoparticles from nucleic acids hybridised with the oligonucleotides. 
     
     
       10. The method according to  claim 1 , wherein the method further comprises a global heating step. 
     
     
       11. The method according to  claim 1 , wherein the method further comprises an annealing step and the annealing temperature is equal to an elongation temperature. 
     
     
       12. The method according to  claim 1 , wherein at any one time during the method only a part of the nanoparticles is heated by excitation. 
     
     
       13. The method according to  claim 1 , wherein the method further comprises a directed movement of the sample relative to an exciting field takes place such that at different times, nanoparticles in different partial volumes of the sample are excited. 
     
     
       14. The method according to  claim 1 , wherein the method further comprises use of a thermolabile DNA polymerase. 
     
     
       15. The method according to  claim 1 , wherein the method further comprises concentration of the amplification product and the concentration of the product of the amplification reaction is determined using test probes.